cruise report of yokosuka-shinkai-6500 dive cruise …...1 cruise report of yokosuka-shinkai-6500...

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Cruise Report of Yokosuka-Shinkai-6500 dive cruise YK09-12 at Japan Trench from Aug. 28 to Sept. 7, 2009

Tadashi Maruyama and Members of YK09-12 cruise

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Contents

Aims of the cruise………………………………………….…………………………………… 3 Member of the cruise………………………………………………………………………… 3 Cruise Diary……………………………………………………………………………………. 5 Wake of R/V Yokosuka during YK09-12 cruise………………………………………….. 10 Diving area of Shinkai 6500 ……………………………………………………………….. 11 Payloads of Shinkai 6500 for dives #1160 and 1161 …………………………………….12 Dive report of Shinkai 6500 dive #1160 ………………………………………………….. 13 Dive report of Shinkai 6500 dive #1161 ………………………………………………….. 17 Some pictures of organisms observed/collected in YK09-12 ………………………….. 21 Preliminary Research reports……………………………………………………………… 23

Yoshida-team …………………………………………………..……………………….. 23 Jimbo-team …………………………………………………………………………...….. 25 Tsuchiya-team …………………………………………………..………………………..26 Miyake-team …………………………………………………………………………….. 28 Ryukyu Universtiy team ……………………………………………………………… 31

Sample lists ……………………………………………………………………………………33

Sample list 1. Calyptogena clam …………………………………………………….. 33 Sample list 2. Sediments and protists ……………………………………………….35 Sample list 3. Invertebrates and trashes……………………… ……………………37 Sample list 4. Invertebrates ...…………………………………………………………38

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Aims of the Cruise

This research cruise, YK09-12, was planned to collect some deep-sea dwelling chemosynthetically symbiotic clams, Calyptogena phaseoliformis and C. fossajaponica, and to collect some deep-sea dwelling protists in the Japan Trench. Some other unique deep-sea dwelling invertebrates, such as cnidarians and ctenophores, were also planned to be collected.

Members of the cruise Scientists Tadashi Maruyama, D. Sc. Principal Investigator of YK09-12 (Marine Biodiversity Research Program, JASMTEC) Masashi Tsuchia, PhD. (JAMSTEC) Takao Yoshida, PhD. (JAMSTEC) Yoshimitsu Nakamura, PhD. (JAMSTEC) Yuki Hongo, Mr. (Tokyo Univ. Marine Sci. and Technol.) Akihiro Tame, Mr. (Marine Works Japan, Co.) Hiroshi Miyake, PhD. (Kitasato Univ.) Mitsuru Jinbo, PhD. (Kitasato Univ.) Haruka Shibata Ms. (Kitasato Univ.) Takuya Mekawa Mr. (Nippon Marine Enterprises, Ltd.) Béatrice Lecroq, PhD (Univ. Geneva) Cathalina Aguilar Hurtado, Ms. (Univ. Ryukyus) Frederic Sinniger, PhD. (Univ. Ryukyus) Takuma Fujii, Mr. (Univ. Ryukyus) Takuya Maekawa (Nippon Marine Enterprises, Ltd.) Shinkai-6500 Operation Team Toshiaki Sakurai, Mr. (Chief Submersible Staff) Kazuhiro Chiba, Mr. (Sub-chief Submersible Staff) Satoshi Ogura, Mr. (Sub-chief Submersible Staff) Keita Matsumoto, Mr. (1st Submersible Staff) Hirofumi Ueki, Mr. (2nd Submersible Staff) Keigo Suzuki, Mr. (2nd Submersible Staff) Yohsuke Chida, Mr. (2nd Submersible Staff)

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Fumitaka Saitoh, Mr. (2nd Submersible Staff) Hitomi Ikeda, Ms. (3rd Submersible Staff) Takuma Ohnishi, Mr. (3rd Submersible Staff) Masaya Katagiri, Mr. (3rd Submersible Staff) Yudai Tayama, Mr. (3rd Submersible Staff) Crews of R/V. Yokosuka Shinya Ryono, Mr. (Captain) Shin-ichi Kusaka, Mr. (Chief Officer) Hiroyuki Kato, Mr. (2nd Officer) Shunsuke Fujii, Mr. (3rd Officer) Toshihiro Kimura, Mr. (Chief Engineer) Kazuhiko Kaneda, Mr. (1st Engineer) Yoshinobu Hiratsuka, Mr. (2nd Engineer) Ichiro Deguchi, Mr. (3rd Engineer) Hirosuke Saitake, Mr. (Chief Radio Operator) Yohei Yamamoto, Mr. (2nd Radio Operator) Yasuyoshi Kyuuki, Mr. (Boat Swain) Yuuki Yoshino, Mr. (Able Seaman) Katumi Shimizu, Mr. (Able Seaman) Shuji Takuno, Mr. (Able Seaman) Nobuyuki Ichikawa, Mr. (Able Seaman) Naoshi Ishizuka, Mr. (Sailor) Daisuke? Yanagitani, Mr. (Sailor) Kazuaki Nakai, Mr. (No. 1. Oiler) Shota Watanabe, Mr. (Oiler) Tomoyuki Hashimoto, Mr. (Oiler) Kazuo Abe, Mr. (Oiler) Takeshi Watanabe, Mr. (Assistant Oiler) Kaoru Takashima, Mr. (Chief Steward) Yoshinobu Hasatani, Mr. (Steward) Kazuhiro Hirayama, Mr. (Steward) Hiroyuki Ohba, Mr. (Steward) Hidetosi Kamata, Mr. (Steward)

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Cruise Diary

Aug. 28th, 2009. Weather: Cloudy with some rain. Sea surface: a little wavy but not rough. We departed the Ohfunato-port at 14:00 on Aug. 28th. But because of a trouble of the engine of the R/V Yokosuka, we repaired the trouble and stayed in the Ohfunato-bay in the night. We had a meeting at 14:30. We introduced each other and the PI explained the main subjects of this research cruise. We set laboratories and made preparations for payload of the Shinkai-6500. Aug. 29th, 2009. At 6:00 we departed from the Ohfunato-bay and headed to the dive point S-1 (40° 30.0N

144° 50.0E). Arrived at the point around 15:30. Preliminary survey of XBT at the site was done. In

the morning, the submersible staffs, gave a lecture of the Shinkai-6500 (6K) to us. They showed us

the inside of the sphere and several other materials like diver’s specially designed suits for the divers.

We saw two sperm-whales and a sea-turtle around noon. We stayed there for a night.

Aug. 30th, 2009. Because a strong wind warning (probably derived from the typhoon#11) was announced by the

weather department, we decided not to dive there and escaped from the Typhoon #11. We headed to

the Mutsu-bay. During the voyage to the Mutsu-bay, the sea was very calm and weather was so nice

but we were miserable. On the way to the Mutsu-bay, we saw many dolphins (16:40-50) and some

whales (17:40-50).

Figure 1. Weather satellite image of the west Pacific and ship track near Tohoku area at 090830.8:05.

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Aug. 31st. 2009. We stayed in the Mutsu-bay. The sea in the bay was calm. In the evening, we had a seminar. Catalina, Frederic and Tad gave talks on the octocoral, zoanthids and diseases in marine mammals, respectively.

Figure 2. Weather satellite image of the west Pacific and ship track at 090831.8:53

Sept. 1st. 2009. We stayed in the Mutsu-bay. Weather was nice and the sea was calm. In the evening, we had a seminar again. Dr. Jinbo and Beatrice gave talks on the lectin in symbioses, and deep-sea foraminifers, respectively.

Figure 3. Weather satellite image of the west Pacific at 090901.10:18.

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Sept. 2nd, 2009. Departed the Mutsu-bay at 8:00. From the broadcast news, the wave at the coast of Tohoku (Northern east part of Japan) was still high (approximately 3 m), though the weather was fine when we departed the Mutsu-bay. We were planning to dive on Sept. 6th. In the weather satellite image, we saw some clouds in the area where we planned to dive. Because the rough sea condition, we did not have the seminar.

Figure 4. Weather satellite image at 090902.8:18. Sept. 3rd, 2009. The weather was fine but the waves were still too high for the dive. We had prepared for the dive but waited the waves calming down. In the southern ocean, a new tropical depression appeared. At 9:10, we decided not to dive today. We headed to the station, S-2 and made the preliminary survey (XBT) there. We will dive at the S-2 and try to find 2 Calyptogena species and the deep-sea foraminifer. If we find them all, we stay there and dive again in the next day. At the seminar, Miyake-san gave a talk on cultivation of the deep-sea animals in the Shin-Enoshima Aquarium. Tsukahara-san told us two tories; one about development of the new awamori by finding the new species of yeast from a fruit, mango, and the genome analysis of Aspergilus fungus of the awamori fermentation. Tsukahara-san brought in two bottles of the awamori, they developed.

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Figure 5. Weather satellite image of the west Pacific (090903.08:18) and the ship track (090903.14:40).

Sept. 4th, 2009. Cloudy and still wave was high. We made the very first dive in this cruise at S-2. The diver was Dr. Miyake. The S-2 was selected because at this point it was expected to collect most biological samples we needed. The diving was started at 9:00. At about 11:00, the submersible arrived at the bottom and collected, Calyptogena phaseoliformis, some zoanthids, the benthic ctenophore, and some trashes (plastic bags etc) to which some octocorals attached. But we could not find the deep-sea foraminifers and C. fossajaponica. We found a sunken log of 2 m long, on which some sessile organismsattached. It was too big to be collected by the submersible. So that we set a marker near the log.

At night, the meeting was canceled and all members concentrated themselves to their studies with the samples. Sept. 5th, 2009 Weather was fine, probably the best day in this cruise but the wave was still high. We dive at the S-2 site again. The diver was M. Tsuchiya-san. We hoped that he could find the deep-sea foraminifer. In the dive, Calytptogena phaseoliformis, octocorals, the deep-sea fish and the trash material were collected but the deep-sea foraminifer and the C. fossajaponica were not found.

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After the dive, we headed to Ohfunato-bay. Sept. 6th, 2009. In the early morning, we arrived at Ohfunato-bay. The weather was cloudy. At 8:00 am, five members, Drs. Jinbo, Miyake, Tsukahara and Sinniger, and Ms. Shibata, left the R/V, Yokosuka. The rest of the members did some experiments and cleaned up the laboratories. In this cruise, we made only two dives but collected C. phaseoliformis, some octocorals and some other samples.

September 7th, 2009.

At 9:00, we arrived at the warf of JAMSTEC in Yokosuka. We left the R/V Yokosuka with our

personal goods. To avoid the typhoon, The R/V left the warf at 11:00. That was the end of the cruise

YK09-12. We thank the captain, crews and the operation team of the Shinkai 6500.

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The wake of R/V Yokosuka during YK09-12 cruise.

Figure 6. Ship track of the R/V Yokosuka in the YK09-12 cruise.

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Diving Area Dive tracks of the dive #1160 and dive #1161 are depicted in Figure 7.

Figure 7. Diving areas of dives #1160 (red) and #1161 (blue).

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Payloads of the Shinaki 6500 for dives # 1160 and 1161. Payload: One suction sampler (multiple canister), one coop sampler,Sample boxes (1 large

and 1 small), 6 MBARI-type cores, 2 MT-type cores, and 2 Markers.

Figure 8. Payload for dive # 1160

Figure Payload for dive # 1161

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Dive report of Shinkai 6500 dive #1160 Dive position: 39° 06.4 N 143°53.5 E

Date：4th September, 2009 Area：5350m Japan Trench, off Miyako, Iwate. Objective：Sampling Calyptogena clams, foraminifera and Zoanthids. Reporter：Hiroshi Miyake (Kitasato University) That was the first dive in YK09-12 after departure from Ofunato port on 28th September, though there was some sea swells by Typhoon. To land at the bottom on the south of the foreordination landing point, we tried to move to the south at the depth of 3500m. However, we descended without moving to south, because there was something wrong with steering of main propeller. Just after landing at a bottom, a colony of Calyptogena phaseoliformis was observed. On the way to approach to the colony, a plastic bag was found and sampled. Then some C. phaseoliformis were collected and MBARI push core (Black) sampling was conducted near the calyptogena colony. During looking for giant foraminifera, Xenophyophor, a sunken wood which was 2 m long was found. Benthic ctenophore was also found near the wood and was collected using MBARI push core (Blue). Some Munidopsis sp. were observed on the sunken wood and collected using slurp gun. A marker #98 was deployed at the sunken wood site. Furthermore looking for giant foraminifera, xenophyophores, a mudstone which was attached many Zoanthid was found. After observing the Zoanthid, sampling was tried, however mudstone was not collected using manipulator of Shinkai 6500 because it was too strong power to grab the mudstone. Then broken mudstone with zoanthids were collected using slurp gun. Next we headed to Event mark #30 to find Calyptogena fossajaponica. During looking for C. fossajaponica, another mudstone with zoanthid was found and sampled using manipulator. Finally, package plastic was collected and two MBARI push core samples were collected at a colony of C. phaseoliformis.

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Events in the dive #1160 recorded by the R/V Yokosuka control room.

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Track of Shinkai 6500 in dive # 1160 on the sea-bottom map..

Figure 8. Dive track of the Shinkai 6500 dive # 1160. Figures on the contour lines indicate

depth in m.

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Dive report of the Shinkai 6500 Dive #1161

Date: Sep. 05, 2009

Site: off Miyako, Japan Trench

Landing: 39-6.2388’N, 143-53.9208’E, 5352 m (11:14)

Leaving: 39-6.4719’N, 143-53.2548’E, 5199 m (15:14)

Main Purpose:

Collection of deep-sea foraminifers

Sub Purposes:

1.Collection of deep-sea clam, Calyptogena phaseoliformis

2.The diversity of deep species of zoanthids and octocorals in the deep sea environment

Payload Equipment:

Suction sampler（多連キャニスター） 1

Scoop sampler 1

Sample box 2 (1 large, 1 small)

MBARI-type core 6

MT-type core 2

Marker 2

Dive Summary

‡ Collected sediment cores with MBARI-type corer from different sites (about 400 m

intervals). The sampling sites locate: 39-6.2388N, 143-53.9207E, 5352 m (core green);

39-6.3011N, 143-53.6437E, 5348 m (core red); 39-6.4496N, 143-53.7117E, 5342 m (core

yellow).

‡ Collected some organisms (fish, gastropod, ctenophore, and rubbish) with suction

sampler or MT-core.

‡ Collected Calyprogena phaseoliformis with scoope sampler into cold insulation box

and with suction sampler.

‡ To find Xenophyophore foraminifers and Calyptogena fossajaponica, we observed wide

areas of our sampling site S-2．

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Events recorded by the R/V Yokosuka control room.

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Track of Shinkai 6500 in dive # 1161 on the sea-bottom map.

Figure 9. Dive track of the Shinkai 6500 dive # 1161. Figures on the contour lines indicate

depth in m.

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Organisms observed and/or collected. Colonies of Calyptogena phaseoliformis were found on the sea floor and some of the clams were collected.

Figure 10. Colonies of Calyptogena phaseoliformis in the dive # 1160.

Fig. 11. Deep-sea ctenophores observed in the dive # 1160.

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Figure 12 . Trash plastic bag on the sea floor.

Figure 13. A sunk log on the sea floor.

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Preliminary Research report Yoshida-team

Symbiosis and Evolution studies of deep-sea invertebrates.

Takao Yoshida (JAMSTEC), Yoshimitsu Nakamura (JAMSTEC), Yuki Hongo (JAMSTEC/ Tokyo University of Marine Science and Technology), Akihiro Tame (Marine works Japan), Masatoshi Tsukahara (Tropical technology center Ltd.),

Tadashi Maruyama (JAMSTEC)

Symbiosis involves interactions between genetically different biological species and plays a key role in evolution of eukaryote to gain new functions. Intracellular symbioses between chemoautotrophic bacteria and marine invertebrates dominate the fauna at deep-sea hydrothermal vents and seeps. The host invertebrates are nutritionally dependent on the intracellular chemoautotrophic bacteria. However, the detailed mechanisms of their symbiosis remain unknown. Reductive genome evolution has occurred in vertically transmitted intracellular symbionts. The detailed process of reductive genome evolution is also unknown. To understand the mechanisms of reductive genome evolution, deep-sea bivalves, Calyptogena phaseoliformis clams were collected from a seep in Japan trench, Japan at a depth of 5300 m during dives (Dive #1160 and #1161) of the submersible “Shinkai 6500” (Table 1). The clams were immediately dissected, and DNA was extracted from the gill tissue, and stored at -20ºC. Other tissues were frozen in liquid nitrogen and stored at -80ºC.

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Table 1 Sample list

On board No. Species Name Locality Site Locality Area

Depth

(m)

Lat

deg Lat min N/S

Long

deg Long min E/W Date

No.of

inds.

Dive/Collecting

Methods Fixation Remarks

YK09-12 6K

#1160 B CP 01

Calyptogena

phaseoriformis S2 Japan Trench 5347 39 06.312 N 143 53.563 E 2009.9.4 1 6K#1160 -80 Experiment

YK09-12 6K

#1160 B CP 02

Calyptogena


YK09-12 6K

#1160 B CP 03

Calyptogena


YK09-12 6K

#1160 B CP 04

Calyptogena


YK09-12 6K

#1160 B CP 05

Calyptogena


YK09-12 6K

#1160 B CP 06

Calyptogena


YK09-12 6K

#1160 B CP 07

Calyptogena


YK09-12 6K

#1160 B CP 08

Calyptogena


YK09-12 6K

#1160 B CP 09

Calyptogena


YK09-12 6K

#1160 B CP 10

Calyptogena


YK09-12 6K

#1160 B CP 11

Calyptogena


YK09-12 6K

#1160 B CP 12

Calyptogena


YK09-12 6K

#1160 B CP 13

Calyptogena


YK09-12 6K

#1160 B CP 14

Calyptogena


YK09-12 6K

#1160 B CP 15

Calyptogena


YK09-12 6K

#1160 B CP 16

Calyptogena

phaseoriformis S2 Japan Trench 5347 39 06.312 N 143 53.563 E 2009.9.4 many 6K#1160 -80 Experiment

YK09-12 6K

#1161 B CP 01

Calyptogena


YK09-12 6K

#1161 B CP 02

Calyptogena


YK09-12 6K

#1161 B CP 03

Calyptogena


YK09-12 6K

#1161 B CP 04

Calyptogena


YK09-12 6K

#1161 B CP 05

Calyptogena


YK09-12 6K

#1161 B CP 06

Calyptogena


YK09-12 6K

#1161 B CP 07

Calyptogena


YK09-12 6K

#1161 B CP 08

Calyptogena


YK09-12 6K

#1161 B CP 09

Calyptogena


YK09-12 6K

#1161 B CP 10

Calyptogena


YK09-12 6K

#1161 B CP 11

Calyptogena


YK09-12 6K

#1161 B CP 12

Calyptogena


YK09-12 6K

#1161 B CP 13

Calyptogena


YK09-12 6K

#1161 B CP 14

Calyptogena


YK09-12 6K

#1161 B CP 15

Calyptogena


YK09-12 6K

#1161 B CP 16

Calyptogena

phaseoriformis S2 Japan Trench 5285 39 06.462 N 143 53.319 E 2009.9.5 many 6K#1161 -80 Experiment

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Jimbo-team Comparison of lectin between Calyptogena okutanii and C. phaseoliformis Mitsuru Jimbo (Kitasato University) Objective: We previously purified a carbohydrate binding protein lectin from Calyptogena okutanii. The lectin named COL was the most active at 4°C, in more than 500 mM NaCl with 10 mM CaCl2, which condition is similar to their habits. COL has affinity to lipopolysaccharide from Escherichea coli. Moreover the lectin agglutinated the symbiotic bacteria separated from host. Thus the lectin involved to symbiosis of symbiotic bacteria, since lectins often binds to cell surface fo cells to stimulate them. Since Calyptogena solidissima also contained a lectin similar to COL in respect to molecular mass and N-terminal amino acid sequence. Thus, the similar lectin to COL may exist in other Calyptogena. Recently, we raised anti-COL antibody from rabbit. if the lectins of Calyptogena was similar each other, the lectins should be detected by this antibody. In this research, I get each tissues from Calyptogena phaseoriformis to examine the similar lectin exists. Method: The blood were drewed by syringe, and centrifuged at 2,500 rpm for 5 min. supernatant was used as haemolymph, and pellets were used as hemocytes. Gill, foot, mantle, and adductor muscle were obtained and store at -80°C. Blood cells were fixed by 4% paraformaldehyde in filtrated sea water. It was washed with FSW, and stored at 4°C. Results: Calyptogena phaseoliformis were obtained at shinkai dive #1160. I used fifteen individuals. Tissue samples (gill, foot, mantle, adductor muscle, blood cells and hemolymph) were obtained from five C. phaseoliformis and stored. Blood were collected from ten C. phaseoliformis, and centrifuged. The supernatant and pellet were stored at -80°C. The supernatant has haemagglutinating activity. From one clam, blood cells were fixed.

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Tsuchia-team Distribution and genetic diversity of Xenophyophoreans and soft-shelled foraminifers at abyssal site in

the northwest Pacific

Tsuchiya, Masashi (BioGeos, JAMSTEC), Lecroq, Béatrice (University of Geneve)

Purpose

This study aims to clarify the genetic diversity and ecological roles of the deep-sea population, especially

of the protistan population including Xenophyophore and soft-shelled foraminifera, by using molecular

techniques and stable isotope measurements to understand the evolution, genetic diversity, and

trophic level of Xenophyophores, their ecological roles in the deep-sea are not clarified. In this cruise,

we will conduct visual observation from the Shinkai 6500 to understand the distribution,

density and ecology of Xenophyophores, and we will also collect samples of

sediment/organisms.

Background

Although protists, particularly foraminifers, form an ecologically important link between

bacteria and macrobenthos in biological and physical cycles in nature, not enough studies

have been conducted to clarify this. In addition, the role of protists in the deep-seas is not

clear. Studies on protists are indispensable to clarify the biological diversity of the deep-sea

floor. Xenophyophore, large unicellular organisms, have (a) large cell up to 10 ~ 15 cm in diameter,

making their body with reticulate, massive, bush or fan-like structures. Several studies have

been conducted for classification and time-lapse observation (Gooday et al., 1993) so far, but

not enough ecological studies have been carried out. Recently, molecular phylogenetic studies

were conducted for soft-shelled foraminifers and Xenophyophores, Xenophyophores are

genetically identified with foraminifers (Pawlowski et al. 2003), closely related to

monothalamous soft-shelled foraminifers. Molecular phylogenetic studies and ultrastructural

observation were carried out for Xenophyophore that we collected at proposed S-3 site during

YK07-15 cruise; Lecroq (2009) identified the novel Xenophophore as Shinkaiya lindsayi. However, the ecology and the role of protists in the deep-seas is not clear.

Research results

We conducted; 1) visual observation of landscape, sediment facies, organisms and sampling, observation

with underwater video and still camera, 2) Sampling of sediment cores, 3) Sampling of organisms. Sediment

core samples, listed in the sample list, will be used for foraminiferal genetic diversity (environmental DNA

analyses) and compare other abyssal sites to understand foraminiferal biodiversity, distributuion and

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dispersal mechanisms. Unfortunately, we could not dive into our proposed area S-3, due to bad weather

conditions; we could not collect Xenophyophorean foraminifers.

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Miyake-team On boad report Taxonomic study of benthic ctenophore Hiroshi Miyake and Haruka Shibata (Kitasato University) Objective I found a benthic ctenophore at a depth of 5350m of Japan Trench in the 6K #959 Dive. At that time, I could not sample this species. Two years ago, in 2007, Dhugal Linday of JAMSTEC tried to sample this species in good condition. Though sampling was succeeded in situ, this specimen was destroyed and melt down in sampling gear when Shinkai 6500 returned on deck. In this cruise, I would like to sample the benthic ctenophore in good condition, to describe the morphology of this species, and to analyze DNA. Preliminary report In the 6K DIVE #1160 and #1161, Two benthic ctenophore were collected. The ctenophore was good condition when it was collected in MBARI push core sampler and MT push core sampler. However these specimens were destroyed and melt down when these sample were in my hand. Pressure decrease or high temperature on surface would destroy these sample. However, Some fragments of benthic ctenophores were obitained and fixed them in 99.5% Et-OH for DNA On the other hand, hydrozoa was found on the shell of snail collected in the 6K #1161 Dive. This species may still alive. Especially medusa buds were observed in the colony of the hydorozoa. Also some stephonoscyphus sp. were collected from the mud-stone. Further study 1. DNA analysis of benthic ctenophores, Stephanoscyphus sp. and hydorozoa 2. Cultivation of hydorozoa

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Study of deep-sea debris distribution and animals on and around them at deep-sea floor at

Japan Trench, off Sanriku

Haruka Shibata and Hiroshi Miyake (Kitasato University)

Introduction

Distribution and composition of marine debris at seafloor have been investigated by

bottom trawl surveys off Iwate (Goto, 2006), in Tokyo Bay (Kuriyama et al, 2003) and Kagosima

Bay (Ohtomi et al, 2004). These surveys revealed that most of the marine debris collected from

seafloor were derived from our daily life. The trawl surveys of marine debris were able to give us the

quantitative data of volume, however, the distribution and condition of them at seafloor in detail was

not unclear. In recent years, many marine debris on deep-sea floor where trawl surveys could not

operate were observed using deep-sea submersibles. Analyzing the occurrence of deep-sea debris

from the video footages of deep-sea dive off Sanriku using submersibles, what was surprising was

that many deep-sea debris such as anchors, iron pipes, plastics, cans, and fishing implements were

observed. Moreover, sea anemones and feather stars attached to these deep-sea debris. The deep-sea

floor where these attached animals were observed was not suitable for their habitat, because where is

not hard bottom but soft muddy bottom. However these deep-sea debris were used as substrate for

these attached animals in the place where they can not inhabit originally.

It was impossible to observe attached organisms on deep-sea debris in detail that occurred

on video image. Therefore it was very difficult to identify these attached organisms and whether

these organisms live at the depth originally or not. It is necessary to collect deep-sea debris from

deep-sea for observing attached organisms in detail. The aim of this cruise is to observe deep-sea

debris in situ and to collect them for observation what kind of organism use deep-sea floor debris as

attached substrate.

Materials and Methods

Some deep-sea debris were collected by the manipulator of Shinkai 6500. These collected

debris were sketched, taken pictures, measured the size, and observed the attached organisms. When

attached organism was found, it was taken pictures and recorded features. After obsevation, Sample

was preserved in deep freezer (-80 °C). After this cruise, deep-sea debris will be weigh and attached

organisms will be identified.

Results

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Four deep-sea debris were collected during this cruise (Table 1)

Table1. Collected deep-sea debris in the cruise YK09-12

Date Dive No. Depth (m) Deep-sea debris Attached organism

2009.09.04 1160 5347 Plastic Foramionifera, Octocoral, Polychaeta

2009.09.04 1160 5360 Plastic bag Foramionifera, Octocoral, Polychaeta

2009.09.04 1160 5360 Disposable diaper Foramionifera, Polychaeta

2009.09.05 1161 5350 Towel Pycnogonida

Future study

We would like to make an analysis of the biological and physico-chemical environments where

deep-sea debris were collected and to identify attached organisms on them.

References

Goto, T, (2006) Comments on marine debris distribution and ghost-fishing on the continental slope

off Iwate Prefecture estimated from bottom trawl survey. Nippon Suisan Gakkaishi 72，501-506.

(in Japanese)

Kuriyama, Y., T. Tokai, K. Tabata and H Kanehiro (2003) Distribution and composition of litter on

seabed of Tokyo Bay and its age analysis. Nippon Suisan Gakkaishi 69，770-781. (in Japanese)

Ohtomi, J., S. Fujieda, M. Higashi and A. Habano (2004) Preliminary Trawl Survey for Estimating

Distribution of benthic animals and marine debris in Kagosima Bay Bull. Jpn. Soc. Fish. Occanogr.

68，158-164. (in Japanese)

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Ryukyu-univ. team

September 5, 2009

Frederic Sinniger, Takuma Fujii and Catalina Aguilar

University of the Ryukyus- MISE laboratory

Report Yokosuka 09-12/ Shinkai- 6500

Purpose:

The metazoan diversity part of this research aimed essentially to understand the diversity of deep

species of zoanthids and octocorals in the deep sea environment of the Japan Trench. Few specimens

have already been collected during the 2007- YK07-15 and additional samples are necessary to

describe those species.

Another purpose of this research cruise was to obtain sediment and various metazoan samples in

order to investigate metazoan biodiversity using massive sequencing and barcoding methods. The

use of a new fixation method (DESS) allowing both molecular and morphological analyses was

tested on a pert of the sediment samples.

Results:

Zoanthids and octocorals were collected by manipulator and slurp gun during the two dives at S2

site.

Two zoanthid polyps were collected during the 1160 dive, the species was identified as underscribe

Abyssoanthus sp. The sample was found on the surface of a muddy rock. One polyp was fixed in

ethanol 80% and the other one was transferred to Dr. Tsuchiya and frozen at -80 ºC .

Six octocorals colonies were collected during the two dives (1160 and 1161), the undescribed

colonies were identified as Stolonifera, genus Clavularia. One of the colonies was found attached to

a plastic bag and the remaining on the surface of muddy rocks. The samples were fixed in ethanol

80% and one in formalin. Subsamples were made for isotope analysis by Dr. Masashi Tsuchiya and

preserved in -80 ºC.

Multiple metazoan or potential metazoan samples were collected from the various samples brought

back to the surface. These samples were fixed in 80% ethanol for further DNA analyses.

32

A few sediment samples were collected and fixed with DESS or frozen at -80 ºC. those sediments

were subsamples from push cores. A part of the sediments were sieved and separated in two fractions

(over 300 micro-m and between 300 and 63 micro-m).

Further analysis:

Unfortunately not enough zoanthid could be sampled to complete the analyses, however, selected

markers (COI, 16S and ITS rDNA) will be sequenced and compared to the samples previously

collected. Octocoral samples will be used for DNA extraction in order to compare with the previous

sample and with other octocoral species; moreover, morphological descriptions will be made to

complete the description of this new species.

The various metazoan samples will be analysed following the barcoding concept, using universal

primer to amplify mitochondrial COI or the nuclear 18S, to identify these organisms. Sediment

samples will be partially analysed using massive DNA sequencing while another part will be

analysed morphologically. These two points aim to investigate the metazoan diversity in the deep

sea.

The diversity of deep sea octocorals and zoanthids being virtually unknown, we will continue to

search for anthozoans species in the deep sea to understand more about their diversity in these

unique habitats.

33

Sample list 1. Calyptogena clams

34

Sample list 1. Continued.

35

Sample list 2. Sediments and protists. On

boardNo.

Species NameJapanease Name

Identified by

LocalitySite

LocalityArea

Depth(m)

Latdeg

Lat min N/SLongdeg

Long min E/W DateNo.ofinds.

Dive/Collecting Methods

Taxon

Code

Presentlocation

JAMSTECNo.

Fixation Preservation Remarks

6K-#1160-C-BL-1

SedimentBeatriceLecroq

S2 (YK09-12)

JapanTrench, off

Sanriku5347 39 6.3296 N 143 53.5633 E 04.09.2009 X

ShinkaiPush Core

GenevaUniversity

RNAlater -80CBlack push core

(disturbed interface +polychaete)

6K-#1160-C-BL-2


S2 (YK09-12)

JapanTrench, off

Sanriku5347 39 6.3296 N 143 53.5633 E 04.09.2009 X

ShinkaiPush Core

GenevaUniversity



6K-#1160-C-BL-3


S2 (YK09-12)

JapanTrench, off

Sanriku5347 39 6.3296 N 143 53.5633 E 04.09.2009 X

ShinkaiPush Core

GenevaUniversity



6K-#1160-C-BL-4


S2 (YK09-12)

JapanTrench, off

Sanriku5360 39 6.4366 N 143 53.4629 E 04.09.2009 X

ShinkaiPush Core

GenevaUniversity

RNAlater -80CYellow push core (+

polychaete)

6K-#1160-C-BL-5


S2 (YK09-12)

JapanTrench, off

Sanriku5360 39 6.4366 N 143 53.4629 E 04.09.2009 X

ShinkaiPush Core

GenevaUniversity


polychaete)

6K-#1160-C-BL-6


S2 (YK09-12)

JapanTrench, off

Sanriku5360 39 6.4366 N 143 53.4629 E 04.09.2009 X

ShinkaiPush Core

GenevaUniversity


polychaete)

6K-#1160-C-BL-7


S2 (YK09-12)

JapanTrench, off

Sanriku5360 39 6.4366 N 143 53.4629 E 04.09.2009 X

ShinkaiPush Core

GenevaUniversity

RNAlater -80C Red push core

6K-#1160-C-BL-8


S2 (YK09-12)

JapanTrench, off

Sanriku5360 39 6.4366 N 143 53.4629 E 04.09.2009 X

ShinkaiPush Core

GenevaUniversity

RNAlater =U24-80C Red push core

6K-#1160-C-BL-9


S2 (YK09-12)

JapanTrench, off

Sanriku5360 39 6.4366 N 143 53.4629 E 04.09.2009 X

ShinkaiPush Core

GenevaUniversity

RNAlater -80C Red push core

6K-#1160-C-BL-10


S2 (YK09-12)

JapanTrench, off

Sanriku5347 39 6.3296 N 143 53.5633 E 04.09.2009 X

ShinkaiPush Core

GenevaUniversity

DNA extractionbuffer

4CBlack push core


6K-#1160-C-BL-11


S2 (YK09-12)

JapanTrench, off

Sanriku5347 39 6.3296 N 143 53.5633 E 04.09.2009 X

ShinkaiPush Core

GenevaUniversity


4CBlack push core


6K-#1160-C-BL-12


S2 (YK09-12)

JapanTrench, off

Sanriku5347 39 6.3296 N 143 53.5633 E 04.09.2009 X

ShinkaiPush Core

GenevaUniversity


4CBlack push core


6K-#1160-C-BL-13


S2 (YK09-12)

JapanTrench, off

Sanriku5360 39 6.4366 N 143 53.4629 E 04.09.2009 X

ShinkaiPush Core

GenevaUniversity


4CYellow push core (+

polychaete)

6K-#1160-C-BL-14


S2 (YK09-12)

JapanTrench, off

Sanriku5360 39 6.4366 N 143 53.4629 E 04.09.2009 X

ShinkaiPush Core

GenevaUniversity



polychaete)

6K-#1160-C-BL-15


S2 (YK09-12)

JapanTrench, off

Sanriku5360 39 6.4366 N 143 53.4629 E 04.09.2009 X

ShinkaiPush Core

GenevaUniversity



polychaete)

6K-#1160-C-BL-16


S2 (YK09-12)

JapanTrench, off

Sanriku5360 39 6.4366 N 143 53.4629 E 04.09.2009 X

ShinkaiPush Core

GenevaUniversity


4C Red push core

6K-#1160-C-BL-17


S2 (YK09-12)

JapanTrench, off

Sanriku5360 39 6.4366 N 143 53.4629 E 04.09.2009 X

ShinkaiPush Core

GenevaUniversity


4C Red push core

6K-#1160-C-BL-18


S2 (YK09-12)

JapanTrench, off

Sanriku5360 39 6.4366 N 143 53.4629 E 04.09.2009 X

ShinkaiPush Core

GenevaUniversity


4C Red push core

6K-#1160-B-BL-19

unidentifiedforaminifera

BeatriceLecroq

S2 (YK09-12)

JapanTrench, off

Sanriku5347 39 5.4000 N 143 53.8000 E 04.09.2009 1 fragment Shinkai arms

GenevaUniversity


4C

Agglutinated flatforaminifera with

stercomata (chain balls)found on plastic bag

6K-#1160-B-BL-20


BeatriceLecroq

S2 (YK09-12)

JapanTrench, off


GenevaUniversity


4C



6K-#1160-B-BL-21


BeatriceLecroq

S2 (YK09-12)

JapanTrench, off

Sanriku5347 39 5.4000 N 143 53.8000 E 04.09.2009 1 Shinkai arms

GenevaUniversity


4C


stercomata (chain balls)found on a baby diper

6K-#1160-B-BL-22


BeatriceLecroq

S2 (YK09-12)

JapanTrench, off


GenevaUniversity

Formaline 4% 4C



6K-#1161-C-BL-23


S2 (YK09-12)

JapanTrench, off

Sanriku5352 39 6.2388 N 143 53.9207 E 05.09.2009 X

ShinkaiPush Core

GenevaUniversity

Frozen -80CSterile, core profile,

Green push core

6K-#1161-C-BL-24


S2 (YK09-12)

JapanTrench, off

Sanriku5352 39 6.2388 N 143 53.9207 E 05.09.2009 X

ShinkaiPush Core

GenevaUniversity


Green push core

6K-#1161-C-BL-25


S2 (YK09-12)

JapanTrench, off

Sanriku5352 39 6.2388 N 143 53.9207 E 05.09.2009 X

ShinkaiPush Core

GenevaUniversity


Green push core

6K-#1161-C-BL-26


S2 (YK09-12)

JapanTrench, off

Sanriku5352 39 6.2388 N 143 53.9207 E 05.09.2009 X

ShinkaiPush Core

GenevaUniversity


Green push core

6K-#1161-C-BL-27


S2 (YK09-12)

JapanTrench, off

Sanriku5352 39 6.2388 N 143 53.9207 E 05.09.2009 X

ShinkaiPush Core

GenevaUniversity


Green push core

6K-#1161-C-BL-28


S2 (YK09-12)

JapanTrench, off

Sanriku5352 39 6.2388 N 143 53.9207 E 05.09.2009 X

ShinkaiPush Core

GenevaUniversity


Green push core

6K-#1161-C-BL-29


S2 (YK09-12)

JapanTrench, off

Sanriku5352 39 6.2388 N 143 53.9207 E 05.09.2009 X

ShinkaiPush Core

GenevaUniversity


Green push core

6K-#1161-C-BL-30


S2 (YK09-12)

JapanTrench, off

Sanriku5352 39 6.2388 N 143 53.9207 E 05.09.2009 X

ShinkaiPush Core

GenevaUniversity


Green push core

6K-#1161-C-BL-31


S2 (YK09-12)

JapanTrench, off

Sanriku5352 39 6.2388 N 143 53.9207 E 05.09.2009 X

ShinkaiPush Core

GenevaUniversity


Green push core

6K-#1161-C-BL-32


S2 (YK09-12)

JapanTrench, off

Sanriku5352 39 6.2388 N 143 53.9207 E 05.09.2009 X

ShinkaiPush Core

GenevaUniversity


Green push core

6K-#1161-C-BL-33


S2 (YK09-12)

JapanTrench, off

Sanriku5342 39 6.4496 N 143 53.7117 E 05.09.2009 X

ShinkaiPush Core

GenevaUniversity


Yellow push core

6K-#1161-C-BL-34


S2 (YK09-12)

JapanTrench, off

Sanriku5342 39 6.4496 N 143 53.7117 E 05.09.2009 X

ShinkaiPush Core

GenevaUniversity


Yellow push core

6K-#1161-C-BL-35


S2 (YK09-12)

JapanTrench, off

Sanriku5342 39 6.4496 N 143 53.7117 E 05.09.2009 X

ShinkaiPush Core

GenevaUniversity


Yellow push core

6K-#1161-C-BL-36


S2 (YK09-12)

JapanTrench, off

Sanriku5342 39 6.4496 N 143 53.7117 E 05.09.2009 X

ShinkaiPush Core

GenevaUniversity


Yellow push core

6K-#1161-C-BL-37


S2 (YK09-12)

JapanTrench, off

Sanriku5342 39 6.4496 N 143 53.7117 E 05.09.2009 X

ShinkaiPush Core

GenevaUniversity


Yellow push core

6K-#1161-C-BL-38


S2 (YK09-12)

JapanTrench, off

Sanriku5342 39 6.4496 N 143 53.7117 E 05.09.2009 X

ShinkaiPush Core

GenevaUniversity


Yellow push core

6K-#1161-C-BL-39


S2 (YK09-12)

JapanTrench, off

Sanriku5342 39 6.4496 N 143 53.7117 E 05.09.2009 X

ShinkaiPush Core

GenevaUniversity


Yellow push core

6K-#1161-C-BL-40


S2 (YK09-12)

JapanTrench, off

Sanriku5342 39 6.4496 N 143 53.7117 E 05.09.2009 X

ShinkaiPush Core

GenevaUniversity


Yellow push core

6K-#1161-C-BL-41


S2 (YK09-12)

JapanTrench, off

Sanriku5342 39 6.4496 N 143 53.7117 E 05.09.2009 X

ShinkaiPush Core

GenevaUniversity


Yellow push core

6K-#1161-C-BL-42


S2 (YK09-12)

JapanTrench, off

Sanriku5342 39 6.4496 N 143 53.7117 E 05.09.2009 X

ShinkaiPush Core

GenevaUniversity


Yellow push core

6K-#1160-C-MT1

Sediment TsuchiyaS2 (YK09-

12)

JapanTrench, off

Sanriku5347 39 6.3296 N 143 53.5633 E 04.09.2009 X MBARI Core JAMSTEC Frozen -80C

Black push core, surfacesediment, 5 ml

6K-#1160-C-MT2


12)

JapanTrench, off


Yellow push core,surface sediment, 5 ml

36

Sample list 2 (continued).

6K-#1160-C-MT3


12)

JapanTrench, off


Red push core, surfacesediment, 5 ml

6K-#1161-C-MT4


12)

JapanTrench, off


Green push core, sievedwith >300µm, 5 ml

6K-#1161-C-MT5


12)

JapanTrench, off


Green push core, sievedwith 125-300 µm, 5 ml

6K-#1161-C-MT6


12)

JapanTrench, off


Green push core, sievedwith 63-125 µm, 5 ml

6K-#1161-C-MT7


12)

JapanTrench, off


Yellow push core, ,sieved with >300µm, 5 ml

6K-#1161-C-MT8


12)

JapanTrench, off


Yellow push core, ,sieved with 125-300 µm,

5 ml6K-

#1161-C-MT9


12)

JapanTrench, off


Yellow push core, ,sieved with 63-125 µm, 5

ml6K-

#1160-C-

Yoshida1


12)

JapanTrench, off

Sanriku5360 39 6.4366 N 143 53.4629 E 05.09.2009 X MBARI Core

JAMSTECHori

(Yoshida,Maruyama)

Frozen -80CGreen core, 0-5cm,

smelled high H2S, H2S @surface=0.987µg/L

6K-#1160-

C-Yoshida

2


12)

JapanTrench, off


JAMSTECHori

(Yoshida,Maruyama)

Frozen -80CGreen core, 6-10cm,smelled high H2S

6K-#1161-

C-Yoshida

3


12)

JapanTrench, off


JAMSTECHori

(Yoshida,Maruyama)

Frozen -80CRed core, 0-5cm, welloxygenated, H2S @

surface=0

6K-#1161-

C-Yoshida

4


12)

JapanTrench, off


JAMSTECHori

(Yoshida,Maruyama)

Frozen -80CRed core, 6-10cm, well

oxygenated

37

Sample list 3. Invertebrates and trashes.

38

Sample list 4. Invertebrates.

On board No. Species NameJapanease Name

Identifiedby

LocalitySite

LocalityArea

Depth(m)

Latdeg

Latmin N/S

Longdeg Long min E/W Date

No.ofinds.

Dive/CollectingMethods

TaxonCode

Presentlocation

JAMSTECNo. Fixation Preservation Remarks

6k-#1160-B-R-#1

Octocorallia/Stolonifera C. Aguilar S2

JapanTrench/ OffSanriku ~5347 39 06.420 N 143 53.5559 E 09/03/2005

6k-#1160/manipulator

U.Ryukyus

Ethanol/formalin/-80 Ethanol Found in rock

6k-#1160-B-R-#2 Sponge F. Sinniger S2

JapanTrench/ OffSanriku ~5347 39 06.356 N 143 53.5013 E 09/03/2005 6k-#1160

U.Ryukyus Ethanol Ethanol

6k-#1160-B-R-#3 Worm T. Fujii S2



6k-#1160-B-R-#4 Eggs T. Fujii S2



6k-#1160-B-R-#5 Anthozoa? F. Sinniger S2



6k-#1160-B-R-#6 Ophiuroidea T. Fujii S2



6k-#1160-B-R-#7 Metazoan? F. Sinniger S2



6k-#1160-B-R-#8



U.Ryukyus Ethanol/-80 Ethanol/-80 Found in rock

6k-#1160-B-R-#9 Metazoan? F. Sinniger S2



6k-#1160-B-R-#10 Worm F. Sinniger S2



6k-#1160-B-R-#11 Worm? F. Sinniger S2



6k-#1160-B-R-#12


JapanTrench/ OffSanriku ~5347 39 06.309 N 143 53.5980 E 09/03/2005

6k-#1160/manipulator


Found inplastic bag

6k-#1160-B-R-#13 Amphipod F. Sinniger S2



6k-#1160-B-R-#14 black sediment F. Sinniger S2


JAMSTEC/Tsuchiya -80

6k-#1160-B-R-#15 Worm T. Fujii S2



6k-#1160-B-R-#16 Sediment from stones F. Sinniger S2

JapanTrench/ OffSanriku ~5347 39 N 143 E 09/03/2005 6k-#1160

U.Ryukyus DESS DESS

6k-#1160-B-R-#17 Zoanthid T. Fujii S2


U.Ryukyus,JAMSTEC/Tsuchiya Ethanol/-80 Ethanol/-80

6k-#1160-B-R-#18 Scyphopolyp T. Fujii S2



6k-#1160-B-R-#19 Scyphopolyp F. Sinniger S2



6k-#1160-B-R-#20 Pogonophore F. Sinniger S2


JAMSTEC/Tsuchiya -80 -80

6k-#1160-B-R-#21

Sediment red corelower F. Sinniger S2


JAMSTEC/Tsuchiya -80 -80

6k-#1160-B-R-#22

Sediment red coresurface F. Sinniger S2

JapanTrench/ Off ~5347 39 N 143 E 09/03/2005 6k-#1160

U.Ryukyus DESS DESS

6k-#1160-B-R-#23

Sediment yellow coresurface F. Sinniger S2

JapanTrench/ Off ~5347 39 N 143 E 09/03/2005 6k-#1160

U.Ryukyus DESS DESS

6k-#1160-B-R-#24 Isopod F. Sinniger S2



6k-#1160-B-R-#25

Blue core deep over300micro-meter F. Sinniger S2


U.Ryukyus DESS DESS

6k-#1160-B-R-#26

blue core deep over63micrometer F. Sinniger S2


U.Ryukyus DESS DESS

6k-#1160-B-R-#27

kurage core deep over300micrometer F. Sinniger S2


U.Ryukyus DESS DESS

6k-#1160-B-R-#28

kurage core deep over63micrometer F. Sinniger S2


U.Ryukyus DESS DESS

6k-#1160-B-R-#29

kurage core deep over63micrometer/2 F. Sinniger S2


U.Ryukyus DESS DESS

6k-#1160-B-R-#30 solenogastre B. Lecroq S2



6k-#1161-B-R-#31




6k-#1161-B-R-#32 Sponge F. Sinniger S2



6k-#1161-B-R-#33 Hydrozoan H. Miyake S2


U.Ryukyus

6k-#1161-B-R-#34 Actiniaria F. Sinniger S2


U.Ryukyus

6k-#1161-B-R-#35




6k-#1161-B-R-#36




6k-#1161-B-R-#37 Brittle Star T. Fujii S2



cruise report of yokosuka-shinkai-6500 dive cruise …...1 cruise report of yokosuka-shinkai-6500...

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